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1.
J Immunol ; 142(12): 4475-82, 1989 Jun 15.
Article in English | MEDLINE | ID: mdl-2498430

ABSTRACT

We have analyzed a series of recombinational events at the IgH chain locus of the B cell lymphoma, NFS-5. Each of these recombinational events results in the replacement of the VH gene segment of the rearranged H chain gene (VhDJh) with that of an upstream germline gene segment. Replacements on the productive and nonproductive alleles have been observed. In each case, the recombination occurs in close proximity to a highly conserved heptameric sequence (5'TACTGTG3') which is located at the 3' end of the VH coding region. In the two examples of recombination on the productive allele that have been analyzed, the initial VHQ52 gene is replaced by different VH7183 genes. On the non-productive allele, sequential replacement events have been analyzed: the initial VHQ52 rearrangement is first replaced by a closely related VHQ52 gene, followed by a second replacement using a VHQ52 pseudogene. Southern blot analysis using VH probes indicates that these recombinations may be accompanied by the deletion of germline VH genes belonging to both the VHQ52 and VH7183 families, suggesting that these gene families are interspersed in the NFS/N mouse.


Subject(s)
B-Lymphocytes/analysis , Gene Rearrangement, B-Lymphocyte , Genes, Immunoglobulin , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Variable Region/genetics , Lymphoma/genetics , Alleles , Animals , Base Sequence , Clone Cells/analysis , Cloning, Molecular , Lymphoma/analysis , Mice , Mice, Inbred BALB C , Molecular Sequence Data
2.
J Immunol ; 142(12): 4483-92, 1989 Jun 15.
Article in English | MEDLINE | ID: mdl-2498431

ABSTRACT

Deletion mapping analysis has shown that members of the VH7183 and VHQ52 gene families are interspersed in the NFS/N mouse. To obtain direct evidence that members of these gene families are physically linked, an NFS/N liver library was constructed and genomic clones were analyzed for hybridization to both VHQ52 and VH7183 gene probes. Four clones were identified which contained both VHQ52 and VH7183 hybridizable restriction fragments. Two clones containing rearranged VHQ52 genes were also found to hybridize with the VH7183 gene probe. Sequence analysis of three of the VH7183-containing restriction fragments indicate that all are pseudogenes which contain interruptions at either the 5' and/or 3' ends of the VH coding region. Given the D-proximal location of at least a portion of the VHQ52 gene family relative to VH7183 in NFS/N mice, and the known correlation between D proximity and the frequency of VH gene utilization, 22 NFS/N-derived pre-B cell lines were analyzed for VHQ52 gene utilization. More than 40% of the identified H chain (VHDJH) rearrangements in this survey used members of this gene family. Furthermore, analysis of poly(A)+ RNA from NFS/N fetal liver and adult spleen also indicates preferential utilization of VHQ52 family in fetal liver. Kinetic studies show, however, that there are no changes in relative utilization throughout fetal ontogeny. The implications of these findings for the expression and randomization of the VH repertoire are discussed.


Subject(s)
Genetic Linkage , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Variable Region/genetics , Multigene Family , Amino Acid Sequence , Animals , B-Lymphocytes/analysis , Base Sequence , Cell Line, Transformed , Cloning, Molecular , Embryonic and Fetal Development , Gene Rearrangement, B-Lymphocyte , Immunoglobulin Heavy Chains/isolation & purification , Immunoglobulin Variable Region/isolation & purification , Kinetics , Liver/metabolism , Mice , Mice, Inbred Strains , Molecular Sequence Data , Spleen/metabolism , Stem Cells/analysis
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